Abstract: BACKGROUND & AIM: To construct the RNA interference vector which can specially induce the expression silence of human DNA repair gene hMGMT. MATERIAL AND METHODS: The hMGMT specific siRNA expression cassette was made by two steps PCR, and then linked with pUC19 to get pU6-MGMTi. Co-transfected with pEGFP-C1 into 16HBE and screened by G418. The expression level of mRNA and protein was detected by RT-PCR and Western Blot respectively. RESULTS: hMGMT specific RNA interfere vector pU6-MGMTi was constructed successfully. Transfected 16HBE cell’s MGMT mRNA and protein expression level were dramatically suppressed. CONCLUSION: MGMT expression silence cell line was built by RNA interfere expression cassette and co-transfection technology, which offered condition for studying the gene function of MGMT.

Key words: hMGMT gene, RNA interfere, 16HBE